Masahiro Matsuzaki

Germin-like protein gene family of a moss, Physcomitrella patens, phylogenetically falls into two characteristic new clades

We identified 77 EST clones encoding germin-like proteins (GLPs) from a moss, Physcomitrella patens in a database search. These Physcomitrella GLPs (PpGLPs) were separated into seven groups based on DNA sequence homology. Phylogenetic analysis showed that these groups were divided into two novel clades clearly distinguishable from higher plant germins and GLPs, named bryophyte subfamilies 1 and 2. PpGLPs belonging to bryophyte subfamilies 1 lacked two cysteines at the conserved positions observed in higher plant germins or GLPs. PpGLPs belonging to bryophyte subfamily 2 contained two cysteines as observed in higher plant germins and GLPs. In bryophyte subfamily 1, 12 amino acids, in which one of two cysteines is included, were deleted between boxes A and B. Further, we determined the genomic structure of all of seven PpGLP genes. The sequences of PpGLPs of bryophyte subfamily 1 contained one or two introns, whereas those of bryophyte subfamily 2 contained no introns. Other GLPs from bryophytes, a liverwort GLP from Marchantia polymorpha, and two moss GLPs from Barbula unguiculata and Ceratodon purpureus also fell into bryophyte subfamily 1 and bryophyte subfamily 2, respectively. No higher plant germins and GLPs were grouped into the bryophyte subfamilies 1 and 2 by our analysis. Moreover, we revealed that PpGLP6 had manganese-containing extracellular superoxide dismutase activity. These results indicated that bryophyte possess characteristic GLPs, which phylogenetically are clearly distinguishable from higher plant GLPs.

In situ photochemical activity of the phytobenthic communities in two Antarctic lakes

Photochemical activity of phytobenthic communities in two freshwater lakes in East Antarctica was estimated using a submersible pulse-amplitude modulation (PAM) chlorophyll fluorometer, to answer the following questions: (1) Are the communities under bright summer photosynthetically active radiation (PAR) photosynthetically active? (2) If active, which community shows the most active signals? (3) Where is the most productive part (or depth) in the lake? Our limnological measurements indicated the two lakes were ultra-oligotrophic. Diving observations revealed that the phytobenthos of the lakes was moss-dominated which had different life-forms (moss shoots in shallow depths of both lakes, moss-pillars in the shallow lake, pinnacle moss-microbial complex community in the deeper lake). In addition, various mat-forming microbial communities inhabited the lake beds. In situ measurements of photochemical parameters indicated that shoots of mosses living just below the littoral slope, and the apical part of the moss pillars, had the highest photosynthetic activity in open water summer conditions, but mat-forming microbial communities and the other moss-microbial complex communities, showed rather lower activity. Most of the mat-forming phytobenthos surface also showed positive photosynthetic activity, but there were some cases of negligible signals in the shallow depth. This suggests that the photosynthetic activities of mat-forming communities in the shallow water were suppressed by strong ambient light in summer.

Differential regulation of periplasmic nitrate reductase gene (napKEFDABC) expression between aerobiosis and anaerobiosis with nitrate in a denitrifying phototroph Rhodobacter sphaeroides f. sp. denitrificans

A denitrifying phototroph, Rhodobacter sphaeroides f. sp. denitrificans, has the ability to denitrify by respiring nitrate. The periplasmic respiratory nitrate reductase (Nap) catalyses the first step in denitrification and is encoded by the genes, napKEFDABC. By assaying the ß-galactosidase activity of napKEFD-lacZ fusions in wild type and nap mutant cells grown under various growth conditions, the environmental signal for inducing nap expression was examined. Under anoxic conditions with nitrate, nap genes expression in the wild-type strain was highest in the dark, and somewhat lowered by incident light, but that of the napA, napB, and napC mutant strains was low, showing that nap expression is dependent on nitrate respiration. Under oxic conditions, both the wild type and nap mutant cells showed high ß-galactosidase activities, comparable to the wild-type grown under anoxic conditions with nitrate. Myxothiazol, a specific inhibitor of the cytochrome bc1 complex, did not affect the ß-galactosidase activity in the wild-type cells grown aerobically, suggesting that the redox state of the quinone pool was not a candidate for the activation signal for aerobic nap expression. These results suggested that the trans-acting regulatory signals for nap expression differ between anoxic and oxic conditions. Deletion analysis showed that the nucleotide sequence from -135 to -88 with respect to the translational start point is essential for nap expression either under anoxic or oxic conditions, suggesting that the same cis-acting element is involved in regulating nap expression under either anoxic with nitrate or oxic conditions.